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Pathogenesis of Bovine Alphaherpesvirus 2 in Calves Following Different Routes of Inoculation1 Bruna P

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Pesq. Vet. Bras. 40(5):360-367, May 2020 DOI: 10.1590/1678-5150-PVB-6588 Original Article Livestock Diseases ISSN 0100-736X (Print) ISSN 1678-5150 (Online) PVB-6588 LD Pathogenesis of Bovine alphaherpesvirus 2 in calves following different routes of inoculation1 Bruna P. Amaral2,3, José C. Jardim4 , Juliana F. Cargnelutti3,5, Mathias Martins6,7 , Rudi Weiblen2,3 and Eduardo F. Flores2,3* ABSTRACT.-Amaral B.P., Jardim J.C., Cargnelutti J.F., Martins M., Weiblen R. & Flores E.F. 2020. Pathogenesis of Bovine alphaherpesvirus 2 in calves inoculated by different routes. Pesquisa Veterinária Brasileira 40(5):360-367. Setor de Virologia, Departamento de Medicina Veterinária Preventiva, Universidade Federal de Santa Maria, Av. Roraima 1000, Rua Z, Prédio 63A, Santa Maria, RS 97105-900, Brazil. E-mail: Título Original Bovine alphaherpesvirus 2 (BoHV-2) is the agent of herpetic mammilitis (BHM), a cutaneous and self-limiting disease affecting the udder [email protected] teats of cows. The pathogenesis of BoHV-2 is pourly understood, hampering the development of therapeutic drugs, vaccines [Título traduzido]. and other control measures. This study investigated the pathogenesis of BoHV-2 in calves after inoculation through different routes. Three- to four-months seronegative calves were 7 -1 inoculated with BoHV-2 (10 TCID50.mL ) intramuscular (IM, n=4), intravenous (IV, n=4) Autores and serological monitoring. Calves inoculated by the IV route presented as light increase orin bodytransdermal temperature (TD) afterbetween mild days scarification 6 to 9 post-inoculation (n=4) and submitted (pi). Virus to virological, inoculation clinical by the by hyperemia, small vesicles, mild exudation and scab formation, between days 2 and 8pi. TDVirus route or viral resulted DNA wasin mild detected inflammatory by PCR in lesions the crusts/swabs at the sites collected of inoculation, from lesions characterized of 3 out of 4 animals inoculated TD from day 2 to 8pi. Viremia was detected in 3/4 animals of the IM group (from day 4 to 8pi); in 2/4 animals of the IV group (days 6 and 8pi) but not in the TD group. Calves from all inoculated groups seroconverted to BoHV-2 in titers from 4 to 64, as indicated by virus-neutralizing (VN) assays performed in sera collected at day 15pi. Administration of dexamethasone (Dex) to the inoculated calves at day 48pi, did not result in virus reactivation as indicated by lack of virus detection in the blood and/or in inoculation sites and no increase in VN antibody titers. These results demonstrated that BoHV-2 was after IM and IV inoculation and was not reactivated by Dex treatment. able to replicate efficiently in calves following different routes of exposure, produced viremia INDEX TERMS: Pathogenesis, Bovine alphaherpesvirus 2, calves, cattle, acute infection, latency. RESUMO.- [Patogênese do alfaherpesvírus bovino 2 em úbere e tetos de vacas. Pouco se sabe sobre a patogênese do bezerros inoculados por diferentes vias.] O alfaherpesvírus bovino 2 (BoHV-2) é um agente etiológico da mamilite terapêuticos e vacinas. Este estudo investigou a patogênese herpética (BHM), uma doença cutânea e autolimitante do doBoHV-2, BoHV-2 dificultando em bezerros o desenvolvimento após a inoculação de medicamentospor diferentes 1 Received on February 19, 2020. Accepted for publication on March 6, 2020. 2 Setor de Virologia, Departamento de Medicina Veterinária Preventiva, 5 Laboratório de Bacteriologia, Departamento de Medicina Veterinária Universidade Federal de Santa Maria (UFSM), Av. Roraima 1000, Prédio Preventiva, Universidade Federal de Santa Maria (UFSM), Av. Roraima 1000, 63A, Santa Maria, RS 97105-900, Brazil. *Corresponding author: Prédio 63C, Santa Maria, RS 97105-900, Brazil. 6 Laboratório de Virologia, Universidade do Oeste de Santa Catarina 3 Graduate Program in Veterinary Medicine (PPGMV), Universidade Federal (Unoesc), Rua Dirceu Giordani 696, Xanxerê, SC 89820-000, Brazil. [email protected] Santa Maria (UFSM), Av. Roraima 1000, Santa Maria, RS 97105-900, Brazil. 7 Department of Population Medicine and Diagnostic Sciences, Animal 4 Laboratório de Virologia, Universidade Federal do Pampa (Unipampa), Health Diagnostic Center, College of Veterinary Medicine, Cornell University, Rodovia BR-472 Km 585, Uruguaiana, RS 97501-970, Brazil. Ithaca, NY 14853, USA. 360 Pathogenesis of Bovine alphaherpesvirus 2 in calves inoculated by different routes 361 vias. Bezerros soronegativos de três a quatro meses foram breed calves in California, US (Watanabe et al. 2017). These inoculados com BoHV-2 (107TCID50.mL-1) por via intramuscular (IM, n=4), por via intravenosa (IV, n=4) ou transdérmica (TD, circulate silently in susceptible host populations, occasionally producingfindings suggest clinical that, cases like to or other outbreaks. simplex viruses,This behavior BoHV-2 maymay virológico, clínico e sorológico. Os bezerros inoculados pela n=4)via IV após apresentaram escarificação aumento leve e submetidosleve da temperatura a monitoramento corporal populations with no evident, or scarce, history of compatible entre os dias 6 a 9 pós-inoculação (pi). A inoculação do vírus clinicalalso explain disease findings (Almeida of positive et al. 2008, BoHV-2 Torres serology et al. 2009b). in cattle In spite of its long story and several decades of scattered inoculação, caracterizadas por hiperemia, pequenas vesículas, studies, several aspects of BoHV-2 biology and pathogenesis pelaexsudação via TD leveresultou e formação em lesões de inflamatóriascrostas, entre leves os dias nos 2 locais e 8pi. de O remain obscure. The pathogenesis of acute and latent vírus ou DNA viral foi detectado por PCR nas crostas/swabs infection remains poorly understood. The few pathogenesis coletados de lesões de 3 de 4 animais inoculados TD do dia studies in cattle were performed two or three decades ago, 2 ao 8pi. Viremia foi detectada em 3/4 dos animais do grupo using old and nowadays obsolete techniques and resources. IM (do dia 4 ao 8pi); em 2/4 animais do grupo IV (dias 6 e Experimental reactivation of the infection has not been 8pi), mas não no grupo TD. Bezerros de todos os grupos consistently demonstrated (Probert & Povey 1975, Turner inoculados soroconverteram o BoHV-2 em títulos de 4 a 64, et al. 1976, Castrucci et al. 1980) and attempts to recover conforme indicado por ensaios de vírus-neutralização (VN) the virus upon explant cultures of sensory nerve ganglia realizados em soro coletado no dia 15pi. Administração de have failed (Letchworth & Carmichael 1982). In some of the dexametasona (Dex) nos bezerros inoculados no dia 48pi, não studies demonstrating latency and reactivation, biases may resultou em reativação do vírus, como indicado pela falta de have been introduced by inoculating the virus intravenously detecção de vírus no sangue e/ou nos locais de inoculação (Castrucci et al. 1972, Scott & Martin 1978). Likewise, the e pela ausência de aumento nos títulos de anticorpos. Estes sites in which the virus may remain latent remain unknown resultados demonstraram que o BoHV-2 foi capaz de replicar (Letchworth & Carmichael 1982). Pathogenesis studies were performed in animal models, with their inherent pitfalls and inoculação, produziu viremia após a inoculação IM e IV e não restrictions (Almeida et al. 2008, Torres et al. 2009a). Thus, a eficientementefoi reativado pelo em tratamento bezerros comseguindo Dex. diferentes vias de better understanding of BoHV-2 pathogenesis in their natural TERMOS DE INDEXAÇÃO: Patogênese, alfaherpesvírus bovino 2, hosts is needed. bezerros, bovinos, infecção aguda, latência. In addition, bovine herpesviruses have been proposed as viral vector for vaccine antigen delivery (Kanekiyo et al. 2019). As such, BoHV-2 may be also a candidate viral vector INTRODUCTION for vaccine use. Therefore, the present study investigated Bovine alphaherpesvirus 2 - formerly bovine herpesvirus type 2 selected aspects of the pathogenesis of BoHV-2 in calves. (BoHV-2) - is an enveloped, double-stranded DNA virus belonging to the family Herpesviridae, subfamily Alphaherpesvirinae, MATERIALS AND METHODS genus Simplexvirus (ICTV 2018). BoHV-2 is genetically and (BoHV-2) antigenically related to Human alphaherpesviruses 1 and Cell culture and virus. Bovine alphaherpesvirus 2 2 (HSV-1, HSV-2) (Sterz et al. 1974, Borchers et al. 1990, strain New York 01 (NY01) was kindly provided by Dr. Fernando A. Ehlers et al. 1999). BoHV-2 is the agent of bovine herpetic Osorio (Department of Veterinary and Biomedical Sciences, University mammillitis (BHM), a vesicular, erosive and necrotic cutaneous of Nebraska, Lincoln, USA). All procedures of virus replication, disease of the udder and teats of cows. The distribution and quantitation and virus neutralizing assays (VN) were performed in prevalence of BoHV-2 infection are poorly understood, but CRIB cells (a MDBK-derived cell line resistant to BVDV). Cells were ® serologic data indicate that the virus is widespread in cattle maintained in Eagle’s minimum essential medium (MEM) (Vitrocell , worldwide (Rweyemamu et al. 1966, Dardiri & Stone 1972, Nova Campinas, São Paulo, Brazil) supplemented with 10% fetal ® Gibbs & Rweyemamu 1977). calf serum (Vitrocell ), penicillin (10,000 IU.mL-1), streptomycin Historically, BoHV-2infection has been described in several ®, countries, including the United States (US) (Dardiri & Stone Darmstadt, Hessen, Germany). The virus inoculum consisted of the 1972), Great Britain (Johnston & Scott 1971), Brazil (Alice supernatant(10mg/mL-1) of and CRIB amphotericin
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    empres watch VOL 29 November 2013 [email protected] | www.fao.org/ag/empres.html Emergence of lumpy skin disease in the Eastern Mediterranean Basin countries Contributors: Sherrylin Wainwright a, Ahmed El Idrissi a, Raffaele Mattioli a, Markos Tibbo b, Felix Njeumi a, Eran Raizman a a Food and Agriculture Organization of the United Nations (FAO); b FAO Regional Office for the Near East (FAO-RNE) Animal Health (OIE). considering the area’s substantial livestock Contents LSD was first reported in Africa, where it population. Turkey maintains over 11.3 million crippled the production potential of cattle and heads of cattle. Iraq, Lebanon and the Syrian 1. Introduction 1 compromised vulnerable livelihoods on the Arab Republic maintain 2.8 million heads, continent. LSD moved beyond Africa in 1989 and the Islamic Republic of Iran maintains 2. Risk assessment 3 when Israel confirmed its first LSD outbreak. for approximately 8.5 million heads. Armenia, In subsequent years Bahrain, Kuwait, Oman, Azerbaijan and Georgia combined maintain 3. Risk management Yemen and the West Bank also reported LSD 4million heads. options for at-risk incursion. Lebanon and Jordan joined LSD- At-risk countries include the Russian affected countries in 2012 and 2013, and Federation to the north and Bulgaria and countries 4 most recently Turkey reported the disease in Greece to the west. This is in line with the 4. Conclusions 5 October 2013 (see Table 1). spread of other significant transboundary There is an imminent risk that LSD will animal diseases (e.g. African swine fever, 5. References 5 (or has already) become endemic in some foot-and-mouth disease, sheep pox and goat countries in the Middle East.